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Updated: Sep 23, 2025
A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells
Published on: May 24, 2011
Primary Macrophage Culture from Porcine Blood and Lungs
Lynnette C Goatley1, Rachel Nash2, Christopher L Netherton3
1African Swine Fever Vaccinology Group, The Pirbright Institute, Pirbright, Woking, UK. lynnette.goatley@pirbright.ac.uk.
Abstract:
Primary cultures represent the most reliable method to isolate and propagate field isolates of African swine fever virus (ASFV ). Within the pig ASFV predominantly targets the reticuloendothelial system for replication; therefore, primary macrophage cell cultures are commonly used to isolate, propagate, and study the virus life cycle in the laboratory. In this chapter we will describe methods for the direct isolation of pulmonary alveolar macrophages by lung lavage and the culture of monocyte-derived macrophages from pig blood. We also include a method for the positive selection of CD14+ monocytes as a source for monocyte-derived macrophages from pig blood using microbeads.
Insights
This study details methods for isolating African swine fever virus (ASFV) using primary pig macrophage cultures. These techniques are crucial for understanding ASFV replication and developing control strategies.
Area of Science:
- Veterinary Virology
- Cell Biology
- Immunology
Background:
- African swine fever virus (ASFV) primarily infects the reticuloendothelial system in pigs.
- Primary cell cultures are essential for isolating and propagating ASFV field isolates.
- Macrophage cell cultures are vital for studying ASFV replication and life cycle.
Purpose of the Study:
- To describe reliable methods for isolating and culturing pig macrophages for ASFV research.
- To provide protocols for obtaining primary macrophages for ASFV propagation and study.
Main Methods:
- Isolation of pulmonary alveolar macrophages via lung lavage.
- Culture of monocyte-derived macrophages from pig blood.
- Positive selection of CD14+ monocytes using microbeads for macrophage derivation.
Main Results:
- Established protocols for obtaining viable primary pig macrophages.
- Demonstrated methods for ASFV isolation and propagation in these cultures.
Conclusions:
- Primary pig macrophage cultures are a reliable system for ASFV isolation and propagation.
- These methods facilitate laboratory studies on ASFV pathogenesis and viral life cycle.
